Method of inhibiting interleukin-1 release

ABSTRACT

Methods useful for inhibiting the release of interleukin-1 and for alleviating interleukin-1 mediated conditions, such as IL-1-mediated inflammation, comprising administration of an effective amount of a pharmaceutically acceptable antioxidant compound such as disulfiram, tetrakis [3-(2,6-di-tert-butyl-4-hydroxyphenyl)propionyloxy methyl]methane or 2,4-di-isobutyl-6-(N,N-dimethylaminomethyl)-phenol.

CROSS REFERENCE TO RELATED APPLICATION

This is a continuation-in-part of Application Ser. No. 026,587, filedMar. 17, 1987, now abandoned.

BACKGROUND OF THE INVENTION

1. Field of the Invention

Interleukin-1 (IL-1) is the name for a family of molecules which havemultiple biological effects. The name interleukin-1 was proposed in1979; and earlier literature reports refer to it by some other name.Murphy, British Journal of Rheumatology, 1985; 24(suppl 1): 6-9, andOppenheim et al., Immunology Today, vol. 2, 45-55(1986). IL-1 issecreted by stimulated macrophages, and has several significantbiological effects, such as mediation of T-lymphocyte proliferation andpyrogenic and proinflammatory effects.

IL-1 activities are summarized in the two above papers. IL-1 has beendescribed to mediate the acute phase response in inflammation, and tohave pyrogenic and proinflammatory effects. IL-1 induces connectivetissue changes, and has been demonstrated to induce the release ofdegradative enzymes from mesenchymal cells that are present at sites ofbony erosion in inflammatory disease states, such as rheumatoidarthritis. Billingham, Brit. J. Rheumatology, 1985: 24(suppl 1): 25-28.Dayer, Brit. J. Rheumatology, 1985: 24(suppl 1): 15-20. The productionof acute phase proteins in the hepatocytes during the acute phase ofinflammation is mediated by IL-1. Whicher, Brit. J. Rheumatology, 1985:24(suppl 1): 21-24.

IL-1 is also involved as a mediator in the inflammatory skin disease,psoriasis. Camp et al., J. Immunology 1986: 137: 3469-3474, and Ristow,Proc. Natl. Acad. Sci. USA 1987: 84: 1940-1944. It is cytotoxic forinsulin producing beta cells in the pancreas, and is thus a causativefactor in the development of diabetes mellitus. Bendtzen et al., Science1986: 232: 1545-1547 and Marx, Science 1988: 239: 257-258. IL-1 alsoappears to be involved in the development of athersoclerotic lesions orathersclerotic plaque. Marx, Science 1988: 239: 257-258. In the absenceor suppression of endogenous prostaglandins, IL-1 stimulates growth andproliferation of vascular smooth muscle cells, which could lead tothickening of vascular walls, such as occurs in atherogenesis. Libby etal., Fed. Proc. Mar. 1, 1987: Vol. 46, no. 3: 975, Abstract 3837.

It would be advantageous to control the release of IL-1, and to be ableto treat IL-1-mediated effects. It would also be advantageous to controlor treat IL-1 mediated inflammations, without production of concomitantside effects known to accompany the use of antiinflammatory steroids andnon-steroidal antiinflammatory agents.

SUMMARY OF THE INVENTION

It has now been found that certain pharmaceutically-acceptable compoundscan be used to inhibit the the release of IL-1, and thus to control ortreat IL-1 mediated conditions. Compounds useful in practicing themethod of the invention include disulfiram, tetrakis[3-(2,6-di-tert-butyl-4-hydroxyphenyl) propionyloxy methyl] methane and2,4-di-isobutyl-6-(N, N-dimethylaminomethyl)-phenol. Although thecompounds have diverse structures, the compounds all have antioxidantactivity. Tetrakis [3-(2,6-di-tert-butyl-4-hydroxyphenyl) propionyloxymethyl] methane (also named as Irganox 1010 or as benzenepropanoic acid:3,5-bis(1,1-dimethylethyl)-4-hydroxy-, tetraester with2,2-bis(hydroxymethyl)1,3-propanediol) is commercially used as ananantioxidant. The causal mechanism of any relationship betweenantioxidant activity of the compounds and their ability to inhibit IL-1release is not known, and the invention is not limited to any particulartheoretical mechanism.

Such compounds can be administered to animals to inhibit secretion ofIL-1; to inhibit or treat IL-1-mediated effects; and to inhibit or treatIL-1-mediated inflammation.

In the method of the invention, one or more compound is administered toan animal, typically to a mammal in need of inhibition of IL-1secretion, inhibition of IL-1-mediated effects, or inhibition of IL-b1-mediated inflammation, in an amount effective to produce suchinhibition. The compounds can be administered to inhibit or treatIL-1-mediated effects in conditions such as inflammation, psoriasis,atherosclerosis, and diabetes.

The compounds can be administered by conventional routes, oraladministration being preferred. The dosage to be employed will varyaccording to factors such as the species, age, weight and condition ofthe particular animal being treated, and the particular compoundemployed. Optimum dosages in particular situations can be determined byconventional dose range finding techniques.

In general, dosage levels for the use of the compounds to inhibit IL-1release can be ascertained by conventional range finding studies. Thecompounds are preferably administered orally at daily dosages from aboutone to about 300 milligrams of active ingredient per kilogram of animalbody weight. Useful results in inhibition of IL-1 release have beenobtained with daily oral dosages of 100 milligrams per kilogram ofanimal body weight (mg/kg).

Although some of the compounds of the method of the invention, such asdisulfiram, are known to produce pharmacologic effects unrelated to IL-1release, they can be usefully employed in the method of the inventionwith animals which are not in need of such other pharmacologic action.In certain cases, the other pharmacologic action may be regarded as anundesirable side effect, when the desired result is inhibition of IL-1release. Thus, with disulfiram, for example, concomitant administrationof ethanol should be avoided, to avoid the well known reaction toalcohol.

The compounds used in the invention can be formulated in conventionalpharmaceutically-acceptable carriers to provide unit dosage formsconvenient for administration. In general, known dosage forms andcarrier materials can be used.

The invention is further illustrated in the following Example.

EXAMPLE

Peritoneal macrophages obtained from CD-1mice were collected and washedonce with RPMI-1640 medium (GIBCO, Grand Island, N.Y.) containing 100Units/ml penicillin, 100 μg/ml streptomycin and 25 μg/ml fungizone(GIBCO, Grand Island, N.Y.). Cells were suspended at 6×10⁶ cells per ml,and one ml aliquots were plated into 6-well flat-bottom plates. Afterone hour incubation at 37° C. in a humidified air chamber containing 5%CO₂, non-adherent cells were removed and 1 ml RPMI medium (with orwithout lipopolysaccharide (LPS) -100 μg/well) was added to each well;LPS stimulates macrophages to release IL-1. Incubation was continued for6 hours, after which the culture supernatant was collected and filteredthrough 0.22 micrometer Acrodisc filters (Gelman, Ann Arbor, MI). Thefluid was stored at a temperature of -70° C. until assayed activity forIL-1 activity.

IL-1 activity was determined by the C3H/HeJ thymocyte proliferationassay of Mizel et al., J. Immunol. 120: 1497 (1978). In this procedure,thymocytes of C3H/HeJ mice are incubated with the macrophage culturesupernatant in the presence of phytohemagglutinin, and pulsed byincubation with ³ H-thymidine. The cells are then harvested andradioactivity is determined by liquid scintillation counting. IL-1activity was expressed as units defined according to Mizel et al, J.Immunol. 120: 1497 (1978).

Compounds were tested in this procedure by oral administration to CD-1mice 40, 24 and 16 hours prior to collection of peritoneal macrophages.The dosage used was 100 mg/kg. The compounds and results obtained areset out in the following table.

    ______________________________________                                                                Percent                                                                       Reduction                                                                     of IL-1                                               Compound                Secretion                                             ______________________________________                                        Disulfiram              79.0%                                                 Tetrakis [3-(2,6-di-tert-butyl-4-                                                                     66.8%                                                 hydroxyphenyl)propionyloxy methyl] methane                                    2,4-Di-isobutyl-6-(N,N--                                                                              92.5%                                                 dimethylaminomethyl)-phenol                                                   ______________________________________                                    

The above results indicate significant inhibition of IL-1 release wasobtained with the test compounds.

What is claimed is:
 1. A method of inhibiting the release ofinterleukin-1 in animals which comprises administering to an animal inneed thereof an amount of disulfiram effective to inhibit the release ofinterleukin-1.
 2. Method of claim 1 wherein the animal is suffering frominflammation and the compound is administered in an amount sufficient toalleviate the inflammation.
 3. Method of claim 1 wherein the animal issuffering from psoriasis and the compound is administered in an amountsufficient to alleviate the psoriasis.
 4. Method of claim 1 wherein theanimal is suffering from diabetes and the compound is administered in anamount sufficient to alleviate the diabetes.
 5. Method of claim 1wherein the animal is suffering from antherosclerosis and the compoundis administered in an amount sufficient to alleviate theatherosclerosis.